EP0213082A1

EP0213082A1 - Substituted hydroxy-heterocyclyl-alkoxy phosphinyloxy trialkylaminium hydroxide inner salt oxides, processes for their preparation and their use as pharmaceuticals

Info

Publication number: EP0213082A1
Application number: EP86810379A
Authority: EP
Inventors: William J. Houlihan; Mark Leonard Lee
Original assignee: Sandoz Erfindungen Verwaltungs GmbH; Sandoz AG
Current assignee: Novartis AG
Priority date: 1985-08-30
Filing date: 1986-08-25
Publication date: 1987-03-04
Also published as: GR862220B; JPH0633293B2; HK74893A; FI83224C; IL79875A; DK414886D0; AU6203886A; IE59077B1; FI863492A; HUT44261A; IL79875A0; AU598744B2; FI863492A0; FI83224B; HU199851B; NZ217389A; EP0213082B1; DK414886A; IE862300L; CY1673A

Abstract

Compounds of formula I

wherein

R represents hydrogen, C _14-18 -alkyl or CH_zOR₂ CH₂CH₂OR₂, wherein R₂ represents C_14-18alkyl
each R, represents independently C_1-3 alkyl m is l or 2
n is an integer from 2 to 8
and R₄ and R₅ together with the adjacent carbon atom represent
wherein R₃ represents alkyl, alkenyl, alkynyl, alkoxy, alkenoxy or alkynoxy each containing 12 to 24 carbon atoms
whereby when R₄ and R₅ together with the adjacent carbon represent (i) ring e) R is H and (ii) other than ring e) R is other than H
which exhibit pharmaceutical in particular anti-tumor activity.

Description

The present invention relates to certain substituted hydroxy-heterocyclylalkoxyphosphinyloxy- trialkylaminium hydroxide inner salt oxides processes for their preparation, pharmaceutical compositions containing them and their use as pharmaceuticals in particular as anti-tumor agents.
More particularly the invention concerns compounds of formula I
wherein

R represents hydrogen, C _14-18 -alkyl or CH₂OR₂ or CH₂CH₂OR₂, wherein R₂ represents C_14-18alkyl
each R, represents independently C_1-3 alkyl m is l or 2
n is an integer from 2 to 8
and R₄ and R_s together with the adjacent carbon atom represent
wherein R, represents alkyl, alkenyl, alkynyl, alkoxy, alkenoxy or alkynoxy each containing 12 to 24 carbon atoms
whereby when R₄ and Rs together with the adjacent carbon represent i) ring e) R is H and ii) other than ring e) R is other than H.

Examples of particular compound groups encompassed within the invention are:

Subclass Ia) wherein R₄ and R_s together with the adjacent carbon atom represent ring a) or b)-,especially ring a);
Subclass Ib) wherein R₄ and R_s together with the adjacent carbon atom represent ring c) or d), especially ring c);
Subclass Ic) wherein R₄ and R_s together with the adjacent carbon atom represent ring g);
Subclass Id) wherein R₄ and R₅ together with the adjacent carbon atom represent ring f).

Within these sub-classes compounds are preferred, wherein R is R' and represents n-C_16-18 alkyl, CH₂OR₂' or CH₂CH₂OR₂' wherein R₂' is n-C₁₆- ₁₈ alkyl and n is an integer from 2 to 6 (sub-classes la' etc). Especially preferred are compounds wherein R is R' ' and represents n-C_16-18 alkyl or CH₂OR₂' and n is an integer from 2 to 4 (sub-classes la ' ' etc). Particularly preferred are compounds wherein R is R' ' as defined above, m is I, n is 2 and each R, is CH₃ (sub-classes la' ' ' etc).
A further subclass of compounds is subclass le wherein R₄ and R_s together with the adjacent carbon atom represent ring e). Within this subclass compounds are preferred wherein R₃ is R₃' and represents alkyl, alkenyl, alkynyl, alkoxy, alkenoxy, alkynoxy each with 12 to 20 carbon atoms m is I and n is an integer from 2 to 6 (subclass le'). Especially, preferred are compounds wherein R₃ is R,' ' and represents alkyl, alkenyl, alkynyl, alkoxy, alkenoxy or alkynoxy each with 12 to 18 carbon atoms, m is I and n is an integer from 2 to 4 (subclass le' '). Particularly preferred are compounds wherein R₃ is R₃''' and represents alkoxy, alkenoxy or alkynoxy, especially alkoxy each with 12 to 24 especially 12 to 18 carbon atoms and each preferably straight chained, m is I and n is an integer from 2 to 4 (subclass le' ' ' ). In the compounds of formula I and in each subclass it is preferred that each R, is the same.
Particularly preferred individual compounds are 2-[hydroxy-(tetrahydro)-2-(octadecyl-5-oxo-2-furanyl)methoxy-phosphinyloxy]N,N,N-trimethylethanaminium hydroxide,inner salt-4-oxide

2-[(2-octadecyloxymethyltetrahydro-2-furanylmethoxy)-hydroxy-phosphinyloxy)-N,N,N-trimethylethanaminium hydroxide, inner salt-4-oxide
2-hydroxy-[I-octadecyloxycarbonyl-3-piperidinyl- methoxy]-phosphinyloxy)-N,N,N-trimethylethanaminium hydroxide, inner salt-4-oxide.

The compounds according to the invention may be prepared by reacting a compound of formula

with a compound of formula CC
wherein R, R,, R₄, R_s, m and n are as defined above and X is a leaving group.
The reaction is suitably carried out in the presence of an inert organic solvent such as a lower alkanol e.g. methanol or ethanol, an aromatic hydrocarbon e.g. toluene or benzene, a dialkylamide such as dimethyl formamide; or acetonitrile at a temperature of e.g. 50° to 70° C. These conditions are particularly suited when employing a preferred leaving group namely bromine, iodine, tosyl or mesyl. The starting material of formula
may be prepared by reacting a compound of formula

with a compound of formula
This reaction is carried out in conventional manner e.g. in the presence of an amine base such as pyridine or triethylamine and in an inert organic solvent, e.g. an aromatic hydrocarbon such as toluene, benzene or xylene, a halogenated aliphatic hydrocarbon, such as methylene chloride, chloroform or carbon tetrachloride, a halogenated aromatic hydrocarbon, e.g. chlorobenzene, or an ether such as diethyl ether. The reaction may be carried out at temperatures of from 20° to 70°C.
The product thus produced is then subjected to basic hydrolysis, e.g., by suspending the product in water. The hydrolysis is conveniently carried out at a temperature of from 20° to 100°C.
The compounds of formula I wherein n is 2 may alternatively be prepared by reacting a compound of formula L
with a compound of formula

wherein R, R,, R₄, R_s and m are as defined above.
The reaction is carried out essentially as described for the reaction of
with
.
The starting materials of formula
, may be prepared by reacting a compound of formula

with a compound of formula
This reaction is carried out in an inert organic solvent such as a halogenated hydrocarbon e.g. methylene chloride, chloroform or an aromatic hydrocarbon e.g. benzene, toluene and in the presence of a tertiary amine such as C_1-4 trialkylamine, e.g. triethylamine and optionally of a catalyst e.g. 4-dimethylaminopyridine at temperatures of e.g. 20_° to 40°C.
The starting material of formula CCC may be prepared according to or analogously to the reaction shown in the following schemes.

LEGEND FOR REACTION SCHEMES

Bn = benzyl
M = alkali or alkaline earth metal
Y = O or S
Z = CH₂ or C=O
E = CI, Br, I or toluenesulfonyl

Examples of conditions for the processes in the reaction schemes are given by way of illustration in the following tables or in the examples hereinafter.

ABBREVIATIONS

10 = inert organic
HC = hydrocarbon
HalHC = halogenated hydrocarbon
THF = tetrahydrofuran
DMSO = dimethylsulfoxide
DMA = dimethylacetamide
DMS = dimethylsulfide
DMF = dimethylformamide

Intermediates containing substituents other than those described in the reaction schemes may be prepared analogously thereto or are either known or may be prepared analogously to known methods.
Starting materials set forth above are either known and/or may be obtained analogously to known methods in conventional manner.
Final products and intermediates may be isolated and purified in conventional manner. Intermediates, where appropriate may be employed directly in the following step without purification.
As is evident to those skilled in the art, the compounds of formula I may exist in racemic or enantiomeric form and the invention is intended to cover all forms.
Enantiomeric forms may be recovered in conventional manner, e.g. by resolution of end or intermediate products or by employing optically active starting materials.
As indicated above, the compounds of formula I are indicated for use as anti-tumor agents and are, therefore, indicated for use in inhibiting the growth of various lymphomas, sarcomas, myelomas and leukemias. The anti-tumor activity of the compounds of formula I may be demonstrated employing the Tumor Cell Cytotoxicity test (TCC test) as follows:
In flat bottom microtiter plates (Nunc Roskilde, Denmark) were placed Abelson 8.1 lymphoma,YAC L1210, P815, Meth A fibrosarcoma or fresh human neuroblastoma tumor cells in DMEM + 10% fetal calf serum and the tumor cell-containing plates were incubated with 1,3 and 5 µg of the test compound for a period of 6 to 72 hours. The number of tumor cells present in the Abelson 8.1, YAC, U210 and P815 assays was determined by measuring the alkaline phosphatase as follows:
The tumor cell plates were centrifuged (500 x g) for ten minutes and the supernatant flicked off. Without further washing, 100 µl of buffer containing 20 µl of diethanolamine, 2nM of MgCl₂. 6H,0, 2.5 µM of p-nitro-phenylphosphate and 10 mg Triton X-100 were added. The samples were incubated for 60 minutes at room temperature and the enzymatic reaction was terminated by the addition of 100 µl of 0.5N NaOH. The absorbance was then measured at 405 nm using a Titertek Multiskan apparatus.
The number of tumor cells present in the Meth A fibrosarcoma and human neuroblastoma assays was measured by ³H-thymidine uptake as follows:
After 72 hours, the cells are thoroughly washed, and each well treated with ca. 0.1µC ³H-thymidine. After 4-6 hours, the cells are collected using a commercial cell harvester, and the radioactivity in the filtrate is measured in a scintillation counter.
The anti-tumor activity of the compounds of formula I may also be demonstrated employing the Influence on Cytotoxicity of ET-18-OCH, test as follows:
Bone marrow cell 'macrophages (10⁵/well) obtained from (BALB/CX57/BL₆)FI mice were incubated with 10µg of I-octadecyl-2-methoxy-3-phosphoryl choline (ET-18-OCH,) for 24 hours in flat bottom microtiter plates (Nunc Roskilde, Denmark), after which time they are centrifuged and washed once. Abelson 8.1 tumor cells in DMEM + 10% fetal calf serum and 1,3 and 5 µg of the test compound were then added to the plates. With the cytotoxicity of ET-18-OCH₃ (10µg) alone set at 100%, the inhibition or enhancement of the cytotoxic effect, as measured by an alkaline phosphatase essay, was determined and values recorded after 72 hours for I, 3 and 5 mg of the test substance.
The usefulness of the compounds of formula I in treating tumors may additionally be demonstrated employing the following procedure:
Meth A fibrosarcoma cells were induced in BALB/C mice by administering methylcholanthrene according to the procedure of Old, et al. (L.J.Old, E.A.Boyse, D.A.Clarke and E.Carswell, Ann.N.Y.Acad.Sci., 101, 80 (1962).
These tumor cells were harvested from the peritoneal cavity 10 to 12 days after administration of methylcholanthrene. Ten CBF, mice of 10-12 week age were each implanted with 7.3 x 10⁶ Meth A sarcoma cells to serve as control. A second group of ten CBF, mice were each implanted with 7.3 x let Meth A sarcoma cells and on day one after implant each mouse was treated p.o. with 5-50µg of the test compound per day for a total of twenty or twenty-seven days. Tumor growth and survivors were assayed on days 7, 14, 21 and 28 after tumor implantation.
The compounds are thus indicated for use as anti-tumor agents and an indicated suitable daily dosage for this use is from about 500 to 2000 mg preferably 1000 to 1500 mg suitably administered in divided dosages of 125 to 2000 mg preferably 250 to 1500 mg one to four times daily or in controlled release form or e.g. 20 mg/kg i.v. over a 24 hr period. A typical oral dosage form contains 300 to 600 mg (unit dose especially 300 to 500 mg in particular 350 to 450 mg).
The invention therefore also concerns a method of treating tumors which comprises administering to a subject in need of such treatment a compound of formula 1, as well as such compounds for use as pharmaceuticals e.g. in treating tumors.
The compounds may be administered alone, or in admixture with a pharmaceutically acceptable diluent or carrier, and, optionally other excipients, and administered orally in such forms as tablets, dispersible powders, granules, elixirs, capsules or suspensions or parenterally in such forms as sterile injectable solutions or suspensions.
The preferred pharmaceutical compositions from the standpoint of ease of preparation and administration are solid compositions, particularly tablets and hard-filled or liquid-filled capsules.
Such compositions also form part of the invention.
The following Examples, in which all temperatures are in °C illustrate the invention.

EXAMPLE I: 2-[hydroxy(tetrahydro)-2-(octadecyloxymethyl-5-oxo-2-furanyl)-methoxyphosphinyloxy]N,N,N-trime-thyl- ethanaminium hydroxide, inner saft-4-oxide

(Subclass Ib;ring c),R = CH₂OC₁₈H₃₇; each R, = CH₃; n = 2, m = 1)

a) Preparation of I-0-octadecyl-3-0-benzylglycerol

To 12.0₉ of 60% sodium hydride in mineral oil - (30 mmol; washed free of oil by the use of petroleum ether) was added 32.4g (30 mmol.) of benzyl alcohol in 200 ml of dry dimethylformamide. The suspension was heated under a flow of nitrogen to 80°C and maintained at this temperature for 45 minutes, after which time 63.2g (0.19 mol) of an epoxide (prepared by the peracid oxidation of al- lylotadecylether) in 100 ml of dimethyl formamide was added and the temperature maintained at 80°C for 15 hours. The solvent was then removed in vacuo and the residue chromatographed on silica gel employing a mixture of petroleum ether and diethyl ether in a ratio of 3:2 as the eluent to yield a low-melting waxy solid.

Preparation of I-O-octadecyl-3-O-benzyldihydrox- yacetone

To a complex prepared at -60°C from the addition of 10.4 ml of dry dimethyl sulfoxide to 8.9 g (70 mmol) of oxalyl chloride in 150 ml of methylene chloride, was added, dropwise, 27.Og - (622 mmol) of the compound prepared in a) above. After stirring the mixture under a nitrogen atmosphere for I hour, 50 ml of triethylamine in 50 ml of methylene chloride was added and the resultant mixture was allowed to warm to 0°C over a period of 30 minutes, after which time it was quenched with 75 ml of water. After the mixture was allowed to warm to room temperature, the organic layer was separated, washed with a saturated sodium chloride solution, dried over magnesium sulfate and concentrated in vacuo to afford an oil which solidified on standing. Flash chromatography on silica gel employing a mixture of petroleum ether and diethyl ether in a ratio of 7:3 as the eluent yielded a white solid.

c) Preparation of 2-benzyloxymethyl-2-hydroxy-l-octadecyloxyhex-5-ene

1.12 g (8 mmol) of 4-bromo-l-butene in 15 ml of dry ether was reacted with 200 mg of magnesium turnings at reflux under a nitrogen atmosphere for I hour. The resulting Grignard reagent was cooled to -60°C and then treated with 2.16 g (5 mmol) of the compound prepared in b) above in 30 ml of ether. After I hour at -60°C, the mixture was warmed overnight to room temperature and then quenched with a saturated aqueous ammonium acetate solution and partitioned. The organic layer was then washed twice with ammonium acetate, washed with a saturated sodium chloride solution and dried over magnesium sulfate, after which time the solvent was removed under reduced pressure. Purification of the crude product was effected on silica gel employing a mixture of petroleum ether and diethyl ether as the eluent to yield a colorless oil.

Preparation of 5-benzyloxymethyl-2-hydroxy-5-oc- tadecyloxymethyltetrahydrofuran

6.0 g (12.3 mmol) of the compound prepared in c) above in 100 ml of methylene chloride was treated with ozone at -60°C. After consumption of the olefin, 25 ml of dimethylsulfide was added and the mixture allowed to warm to room temperature. The solvent was removed in vacuo and the crude product was purified on silica gel employing a mixture of petroleum ether and diethylether in a ratio of 7:3 as the eluent to yield a colorless oil.

e)Preparation of 5-benzyloxymethyl-5-octadecyloxymethyl-4,5-dihydro-2(3H)furanone

To a solution of 1.6g (7.4 mmol) of pyridinium chlorochromate in 25 ml of methylene chloride under a nitrogen atmosphere, was added 132g (6.5 mmol) of the compound prepared in d) above in 25 ml of methylene chloride. After 18 hours at 25°C, the solution was diluted with 150 ml of ether, filtered through silica gel and the filtrate evaporated to afford an oil. The oil was then flash chromatographed on silica gel employing a mixture of petroleum ether and diethyl ether in a ratio of 7:3 as the eluent to yield a colorless oil.

f) Preparation of 5-hydroxymethyl-5-octadecyloxymethyl-4,5-dihydro-2(3H) furanone

A mixture containing 10.7g (21.83 mmol) of the compound prepared in e) above, 300 ml of a mixture of ethyl alcohol and water in a ratio of 9:1 and 1.5g of 5% palladium on carbon (50% water content) was placed in .a pressure bottle and hydrogenated at 40°C under a pressure of 50 lbs. of hydrogen until uptake was complete. The catalyst was then filtered off and the filtrate concentrated in vacuo. The residue was crystallized from methanol to yield a solid.

g) Preparation of the title compound

To 860 mg (2.16 mmol) of the compound prepared in f) above, 29 mg (0.24 mmol) of 4-dimethylaminopyridine and 0.42 ml (3 mmol) of triethylamine in 10 ml of benzene, was added 455 mg (3.2 mmol) of 2-chloro-2-oxo-1,3,2-diox- aphospholane. The resultant mixture was allowed to stir at room temperature for 18 hours, after which time it was filtered and the filtrate was evaporated to dryness. The residue was taken up with 10 ml of dry acetonitrile, cooled to allow introduction of excess condensed trimethylamine, and then heated in a sealed tube under pressure for 20 hours at 70°C. The reaction mixture was then cooled to room temperature, diluted with an equal volume of acetonitrile and filtered to afford a white solid which was recrystallized from a mixture of methylene chloride and acetone to yield a solid, m.p. 215°-220°C - (dec.).

EXAMPLE 2: 2-(hydroxy(tetrahydro)-2-(octadecyl-5-oxo-2-furanyl)methoxyphosphinyloxy]-N,N,N-trimethylethanaminium hydroxide, inner salt -4-oxide

(Subclass Ib; ring c),R = C₁₈H₃₇;m = l, n = 2, each R, = CH,) analogous to example I m.p. 235° -240°C.

EXAMPLE 3: 2-[hydroxy-[tetrahydro-2-(2-octadecyl- oxyethyl)-5-oxo-2-furanyl]-methoxy-phosphinyloxy)-N,N,N-trimethyl ethanaminium hydroxide, inner salt -4-oxide

(Subclass Ib; ring c),R = CH₂CH₂OC₁₂H₃₇; m = I, n = 2, each R, = CH₃) analogous to example I m. p. 107° (dec.)

EXAMPLE 4: 2-[hydroxy-2-[tetrahydro-2-(octadecyl- oxymethyl)-5-oxo-2-furanyl]-ethoxy-phosphinyloxy]-N,N,N-trimethyl ethanaminium hydroxide, inner salt -4-oxide

(Subclass Ib; ring c), R = CH₂OC₁₈H₃₇; m = 2, n = 2, each R, = CH₃) analogous to example I m.p. 145°C (dec.)
EXAMPLE 5: 2-[(2-octadecyloxymethyltetrahydro-2-furanylme-thoxy)-hydroxyphosphinyl-oxy]-N,N,N-trimethylethanaminium hydroxide inner salt -4-oxide
(Subclass la; ring a), R = CH₂OC₁₈H₃₇, m = I,n = 2,each R, = CH₃)

a) Preparation of tetrahydro-2-octadecyloxymethyl-2-furanmethanol

To a mixture of 2.64g (0.020 mol) of 2,2-bis-(hydroxymethyl)-tetrahydrofuran and 2.2g (0.0066 mol) of bromooctadecane in 8 ml of 1:1 mixture of dimethylsulfoxide and tetrahydrofuran was added 1.84 g (0.0264 mol) of finely powdered potassium hydroxide. The resultant mixture was then stirred at room temperature for 2 hours, after which time it was poured onto 100 ml of water, diluted with 20 ml of an aqueous saturated sodium chloride solution and extracted with ether. The ether extract was then washed with an aqueous saturated sodium chloride solution, dried over anhydrous sodium sulfate and the ether was removed under reduced pressure. The crude product was then purified by column chromatography on silica gel employing a mixture of methyl-t-butyl ether and hexane (3:1 ratio) as the eluent to yield the desired compound as a wax.
The title compound is prepared analogously to the last step of example I,white solid m.p. 232-235°C.
EXAMPLE 6: 2-[hydroxy-(tetrahydro)-3-(octadecyl-5-oxo-3-furanyl)-methoxyphosphinyloxy]-N,N,N-trimethyl ethanaminium hydroxide, inner salt -4- oxide
(Subclass Ic; R= C₁₈H₃₇, m = I, n = 2, each R, = CH₃)

a) Preparation of 2-benzyloxymethyl-I-eicosene

To a stirred suspension of 5.0g of methyl triphenylphosphonium bromide in 50 ml of anhydrous tetrahydrofuran and 4.0 ml of hexamethyl- phosphorous triamide was gradually added 27 ml of 0.5M potassium bis-trimethylsilylamide. The resultant mixture was then stirred at room temperature for 30 minutes, after which time it was cooled to -60°C (dry ice/acetone bath) and treated dropwise with a solution of 8.0g (210 mmol) of the compound of Example 2b) in 50 ml of anhydrous tetrahydrofuran and stirred for I hour at -60°C and for I hour at ice bath temperature. The solids were then filtered off and the filtrate evaporated in vacuo to afford an oil. The oil was then chromatographed on silica gel employing a mixture of petroleum ether and methyl-l-butyl ether in a ratio of 9:1 as the eluent to yield a yellow oil.

b) Preparation of 3-benzyloxymethyl-3-octadecyl cyclobutanone

A stirred mixture containing 7.18g (18 mmol) of the compound prepared in a) above, 75 ml of anhydrous diethyl ether and 1.4g of zinc-copper couple was blanketed with nitrogen and treated dropwise at room temperature with a solution containing 3.63 g (20 mmol) of trichloroacetyl chloride and 3.06g (20 mmol) of phosphorus oxychloride. After stirring the mixture at room temperature for 48 hours, the salts were filtered off and the filtrate concentrated in vacuo. The residue was then chromatographed on silica gel employing a mixture of petroleum ether and methyl-t-butyl ether in a ratio of 9:1 as the eluent to yield a yellow oil.

c) Preparation of 4-benzyloxymethyl-4-octadecyl-4,5-dihydro-2(3H)-furanone

A stirred solution containing 2.27g (5.1 mmol) of the-compound prepared in b) above and 22g (12.75 mmol) of m-chloroperbenzoic acid in 30 ml of anhydrous methylene chloride was refluxed for 10 hours under a nitrogen atmosphere. The reaction mixture was then cooled to room temperature, washed with 10 ml of a 2M sodium bisulfite solution, washed twice with 10 ml of a 2M sodium bicarbonate solution and washed with 10 ml of brine, after which time it was dried over magnesium sulfate, filtered and evaporated to afford a colorless liquid. The liquid was then chromatographed on silica gel employing a mixture of petroleum ether and methyl-t-butyl ether in a ratio of 7:3 as the eluent to yield a colorless liquid.
Preparation is continued analogously to example If) and Ig) to produce the title compound as a white solid, m.p. 251°-256°C.

EXAMPLE 7: 2-[(3-octadecyltetrahydro-3-furanyl- methoxy)hydroxyphosphinyloxy]-N,N,N-trimethyl ethanaminium hydroxide, inner salt -4-oxide

(Subclass Id; R = C₁₈H₃₇; m = 1; n = 2, each R, = CH,)

a) Preparation of 3-benzyloxy-3-octadecyl tetrahydrofuran

A solution of 1.0g (2.1 mmol) of the compound of Example 6c) in 25 ml of anhydrous toluene under a nitrogen atmosphere was cooled to -70°C - (methanol/dry ice bath) and treated dropwise, over a period of 45 minutes, with 32 ml of 1.0M diisobutyl aluminium hydride. The resultant mixture was then stirred for an addtional 2 hours, while the temperature was maintained at -70°C, after which time it was carefully poured onto a mixture of I ml of acetic acid and 80g of ice/water. The organic layer was then separated and the aqueous layer extracted twice with 20 ml of toluene The combined toluene layers were then washed with 20 ml of a IN sodium bicarbonate solution and washed twice with 20 ml of brine, after which time they were dried over magnesium sulfate, filtered and evaporated to an oil. The oil was then dissolved in a solution containing 0.366g (3.15 mmol) of triethylsilane and 10 ml of dry methylene chloride, cooled to -20°C (isopropanol/dry ice bath) and treated with 0.326g (2.3 mmol) of boron trifluoride etherate. The resultant mixture was then stirred for I hour, while the temperature was maintained at -20°C, after which time 5 ml of a IN sodium bicarbonate solution was added and the mixture allowed to warm to room temperature. The organic layer was then separated, washed with 5 ml of brine, dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo to afford an oil. The oil was then chromatographed on silica gel employing a mixture of petroleum ether and methyl-t-ether in a ratio of 9:1 as the eluent to yield an oil.
Preparation is continued analogously to example If) and lg) to yield the title compound as a white solid m.p.235°-241°C (dec.)

EXAMPLE 8: 2-[hydroxy(tetrahydro)-3-(octadecyloxymethyl-5-oxo-3-furanyl) methoxyphosphinyloxy]-N,N,N-trimethyl ethanaminium hydroxide, inner salt -4-oxide

(Subclass Ic); R = CH₂OC₁₈H₃₇, m = 1, n = 2, each R, = CH3) analogous to examples 6a), b), c), If), lg); white solid m.p. 220° -225° dec.

EXAMPLE 9: 2-hydroxy-[I-octadecyloxycarbonyl-3-piperidinyl methoxy]-phosphinyloxy]-N,N,N-trimethylethan -aminium hydroxide inner salt -4- oxide

(Subclass le); R₃ = OC₁₈H₃₇; m = I; n = 2; each R, = CH₃)

a) Preparation of 3-hydroxymethyl-N-octadecylox- ycarbonyl piperidine

To a solution of 10.52g (91 mmol) of 3-piperidinemethanol in 100 ml of dichloromethane, and 13.3 ml (96 mmol) of triethylamine was added, at room temperature, a solution of 87 mmol of octadecylchloroformate in toluene. The resultant mixture was then allowed to react for 24 hours, after which time it was washed with water. The aqueous layer was then extracted twice with dichloromethane and the organic layers were combined, dried with magnesium sulfate and filtered. The solvent was then removed in vacuoto yield the crude product as a solid. The crude product was then chromatographed on silica gel employing dichloromethane as the eluent to yield a white solid.

Preparation of the title compound

0.5g (1.22 mmol) of the compound prepared in a) above was dissolved in 20 ml of benzene containing 15mg of N,N-dimethylaminopyridine and 0.21 ml (1.52 mmol) of triethylamine. To the mixture was added 0.23g (1.62 mmol) of 2-chloro-2-oxo-1,3,2-dioxaphospholane and the resultant mixture was stirred at room temperature for 2 hours. The salts were then removed by filtration and the residue taken up in dry acetonitrile which was then cooled to -78°C. Trimethylamine was then condensed therein, the reaction vessel was capped, warmed to 60°C and maintained at this temperature for 24 hours. The reaction mixture was then cooled in an ice-methanol bath and the solids were then isolated by filtration, washed with acetonitrile and dried under vacuum to yield the crude product. The crude product was then chromatographed on silica gel employing a mixture of chloroform, methanol and water (in a 2:1:0.2 ratio) as the eluent to yield a white solid, m.p., >230°C.

EXAMPLE 10: 2-hydroxy-[I-hexadecyloxycarbonyl-3-piperidinylmethoxy]-phosphinyloxy]-N,N,N-trimethylethaneaminium hydroxide inner salt -4-oxide

(Subclass le; R3 = OC₁₆H₃₃, m = I, n = 2, each R, = CH3) analogous to example 9 ,white solid m.p. > 230°.

EXAMPLE II: 2-hydroxy-[I-hexadecanoyl-3-piperidinyl methoxy]-phosphinyloxy]-N,N,N-trimethylethanaminium hydroxide inner salt -4-oxide

(Subclass le; R₃ = C₁₅H₃₁, m = I, n = 2, each R, = CH3) analogous to example 9, white solid; m.p. 229°.

EXAMPLE 12: 6-hydroxy-[I-octadecyloxycarbonyl-3-piperidinyl methoxy]-phosphinyloxy]-N,N,N-trimethylhexanaminium hydroxide inner salt -8-oxide

(Subclass le; R₃ = OC₁₈H₃₇, m = I, n = 6, each R, = CH₃)
To 10 ml of dry benzene containing 0.358g (1.2 mmol) of 6-bromohexyloxyphosphorodichloridate was added 0.411g (1.0 mmol) of the compound of Example la) and the resultant mixture was cooled in an ice-salt bath, after which time it was treated dropwise, under a nitrogen atmosphere, with a solution of 103µl of pyridine in I ml of benzene.
The ice-salt bath was then removed and the mixture allowed to warm to room temperature over a period of 6 hours. The volatiles were then removed under reduced pressure and the residue was suspended in 15 ml of water and heated in the steam bath for I hour. The mixture was then cooled to room temperature, extracted with chloroform and the extracts dried over magnesium sulfate, filtered and the solvent removed in vacuo. The residue was then treated with trimethylamine in a manner analogous to that described above in the last step in the preparation of Example I to afford the hydrobromide salt of the title compound. The hydrobromide salt was then taken up in 10 ml of methanol into which 0.310g of silver carbonate was suspended. After 90 minutes, the solids were removed by filtration and the filtrate concentrated in vacuoto yield the crude product. The crude product was then chromatographed on silica gel employing a mixture of chloroform, methanol and water (in a 2:1:0.2 ratio) as the eluent to yield a white solid, m.p. 137°-140°C.

Claims

I. Compounds of formula I
wherein
R represents hydrogen, C _14-18 -alkyl or CH₂OR₂ or CH₂CH₂OR₂, wherein R₂ represents C_14-18alkyl

each R, represents independently C_1-3 alkyl m is I or 2

n is an integer from 2 to 8

and R₄ and R_s together with the adjacent carbon atom represent

wherein R₃ represents alkyl, alkenyl, alkynyl, alkoxy, alkenoxy or alkynoxy each containing 12 to 24 carbon atoms

whereby when R4 and R_s together with the adjacent carbon represent (i) ring e) R is H and (ii) other than ring e) R is other than H.
2. A compound according to claim I wherein each R, is the same and R₄ and R₅ together with the adjacent carbon atom represent ring a); b); c) or d).
3. A compound according to claim I wherein each R, is the same and R₄ and R_s together with the adjacent carbon atom represent ring e) and m is I.
4. A compound according to claim I selected from those wherein each R, is the same and R₄ and R_s together with the adjacent carbon atom represent
(i) ring a) or b),

(ii) ring a),

(iii) ring c) or d),

(iv) ring c),

(v) ring g),

(vi) ring f) or

(vii) ring e).
5. A compound according to claim 4 selected from those wherein R₄ and R_s together with the adjacent carbon atom have the meanings given in claim 4 (i) to (vi) and
i) R is R' and represents n-C_16-18alkyl, CH₂OR₂' or CH₂CHOR₂' wherein R₂' is n-C_16-18 alkyl and n is an integer from 2 to 6

ii) R is R' ' and represents n-C_16-18 alkyl or CH₂OR₂' wherein R₂' is as defined under (i) and n is an integer from 2 to 4,

iii) R is as defined under (ii), m is I, n is 2 and each R, is CH₃; or

R₄ and R_s together with the adjacent carbon atom represent ring e) and

i) R₃ is R₃' and represents alkyl, alkenyl, alkynyl, alkoxy, alkenoxy, alkynoxy each with 12 to 20 carbon atoms,m is I and n is an integer from 2 to 6,

ii) R₃ is R3' ' and represents alkyl, alkenyl, alkynyl, alkoxy, alkenoxy, or alkynoxy each with 12 to 18 carbon atoms, m is I and n is an integer from 2 to 4,

iii) R₃ is R₃''' and represents alkoxy, alkenoxy, or alkynoxy, especially alkoxy each with 12 to 18 carbon atoms, m is I and n is an integer from 2 to 4.
6. A compound selected from 2-[hydroxy-(tetrahydro)-2-(octadecy)-5-oxo-2-furanyl)methoxy- phosphinyloxy]-N,N,N-trimehtyl-ethanaminium hydroxide, inner salt-4-oxide, 2-[(2-octadecyloxymethyltetrahydro-2-furanylmethoxy)-hydroxyphosphinyloxy]-N,N,N-trimethylethanaminium hydroxide, inner salt-4-oxide.
7. 2-hydroxy-[I-octadecyloxycarbonyl-3-piperidinyl-methoxy]-phosphinyloxy]-N,N,N-trimethylethanaminium hydroxide, inner salt-4-oxide.
8. A pharmaceutical composition comprising a compound according to claim I together with a pharmaceutically acceptable diluent or carrier.
9. A compound according to claim I for use as a pharmaceutical.
10. A compound according to claim I for use as an anti-tumor agent.

II. A process for preparing a compound according to claim I which comprises

a) reacting a compound of formula
with a compound of formula

wherein R, R,, R_«, R_s, m and n are as defined above and X is a leaving group, or

b) when n is 2 reacting a compound of formula

with a compound of formula C C
wherein R, R₁, R₄, R₅ and m are as defined above.